Research Proposal Summary Hepatocellular carcinoma (HCC) is a highly lethal malignancy, and is a worldwide public health problem. The main cancer-related cause of death in patients with HCC is metastatic tumor progression. But the precise molecular mechanisms of HCC metastasis are not well understood. Also, there are no accurate prognostic plasma biomarkers which can help in early detection of aggressive HCC and thereby predict clinical outcome or guide treatment selection. We have developed an autochthonous mouse model of metastatic HCC where conditional overexpression of MYC and Twist1 genes in the mouse liver leads to step-wise progression of liver cancer simulating human HCC. This study proposes to use this novel mouse model of metastatic HCC to explore the mechanisms of tumor progression and identify proteomic biomarkers for HCC. Our central HYPOTHESIS is that Twist1 modulates innate immune system specific proteomic and glycoproteomic changes which promote metastatic progression. In Aim 1 we will perform proteomic and glycoproteomic analyses of MYC HCC and MYC/Twist1 HCC using mass spectrometry. We will then undertake a systems biology approach to large scale proteomics data and perform gene knockout experiments to gain mechanistic insights into how Twist1 promotes metastasis. In Aim 2 we will identify and validate plasma proteomic and glycoproteomic biomarkers of metastatic progression in the MYC/Twist1 mice that are relevant to human HCC. Successful completion of this study has significant translational potential by identifying prognostic biomarkers which will improve treatment allocation. Candidate Career Development Plan The objective of this K08 Mentored Award application is to enable Dr. Renumathy Dhanasekaran to undertake supervised research and career development training to become an independent physician scientist. She is a board certified Hepatologist who joined the Department of Medicine at Stanford in 2015 and is pursuing advanced translational research training under the mentorship of Dr. Dean Felsher. Her long term career goal is to build an independent translational research program to identify biomarkers for HCC. This K08 will provide her with the protected time and support needed to undertake the following training: (1) receive hands-on training in advanced techniques in cancer biology (2) acquire skills in computational biology to perform integrated analyses of genomic data; (3) acquire critical talents required for success in academic medicine like grantsmanship, and communication skills and (4) gather preliminary data required to apply for a NIH R01 grant during year 4-5 of the K08 award. The candidate has an excellent mentor with vast expertise in cancer biology and is working in a well- funded lab. The Chief of the Division of Gastroenterology, will also serve as her co-mentor and he is personally committed to her academic success. She has assembled a team of seasoned mentors with expertise in cancer biology, advanced proteomics, biomedical informatics and clinical hepatology, who will serve on her mentorship advisory committee. Stanford is a world-class institution with particular strength in cancer biology, computational bioinformatics and translational medicine, and hence is the ideal environment to advance her academic career.